Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism

Abstract Promyelocytic leukemia protein (PML) nuclear bodies (NBs) are essential in regulating tumor suppression, antiviral response, inflammation, metabolism, aging, and other important life processes. The re-assembly of PML NBs might lead to an ~100% cure of acute promyelocytic leukemia. However,...

Full description

Saved in:
Bibliographic Details
Main Authors: Yangxia Tan, Jiawei Li, Shiyan Zhang, Yonglei Zhang, Zhiyi Zhuo, Xiaodan Ma, Yue Yin, Yanling Jiang, Yao Cong, Guoyu Meng
Format: Article
Language:English
Published: Nature Publishing Group 2024-11-01
Series:Cell Discovery
Online Access:https://doi.org/10.1038/s41421-024-00735-3
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1846148120210046976
author Yangxia Tan
Jiawei Li
Shiyan Zhang
Yonglei Zhang
Zhiyi Zhuo
Xiaodan Ma
Yue Yin
Yanling Jiang
Yao Cong
Guoyu Meng
author_facet Yangxia Tan
Jiawei Li
Shiyan Zhang
Yonglei Zhang
Zhiyi Zhuo
Xiaodan Ma
Yue Yin
Yanling Jiang
Yao Cong
Guoyu Meng
author_sort Yangxia Tan
collection DOAJ
description Abstract Promyelocytic leukemia protein (PML) nuclear bodies (NBs) are essential in regulating tumor suppression, antiviral response, inflammation, metabolism, aging, and other important life processes. The re-assembly of PML NBs might lead to an ~100% cure of acute promyelocytic leukemia. However, until now, the molecular mechanism underpinning PML NB biogenesis remains elusive due to the lack of structural information. In this study, we present the cryo-electron microscopy (cryo-EM) structure of the PML dimer at an overall resolution of 5.3 Å, encompassing the RING, B-box1/2 and part of the coiled-coil (RBCC) domains. The integrated approach, combining crosslinking and mass spectrometry (XL-MS) and functional analyses, enabled us to observe a unique folding event within the RBCC domains. The RING and B-box1/2 domains fold around the α3 helix, and the α6 helix serves as a pivotal interface for PML dimerization. More importantly, further characterizations of the cryo-EM structure in conjugation with AlphaFold2 prediction, XL-MS, and NB formation assays, help unveil an unprecedented octopus-like mechanism in NB assembly, wherein each CC helix of a PML dimer (PML dimer A) interacts with a CC helix from a neighboring PML dimer (PML dimer B) in an anti-parallel configuration, ultimately leading to the formation of a 2 µm membrane-less subcellular organelle.
format Article
id doaj-art-981e3fbdb2fd4aa29ff0f26ea666ac65
institution Kabale University
issn 2056-5968
language English
publishDate 2024-11-01
publisher Nature Publishing Group
record_format Article
series Cell Discovery
spelling doaj-art-981e3fbdb2fd4aa29ff0f26ea666ac652024-12-01T12:09:24ZengNature Publishing GroupCell Discovery2056-59682024-11-0110111110.1038/s41421-024-00735-3Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanismYangxia Tan0Jiawei Li1Shiyan Zhang2Yonglei Zhang3Zhiyi Zhuo4Xiaodan Ma5Yue Yin6Yanling Jiang7Yao Cong8Guoyu Meng9Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityKey Laboratory of RNA Innovation, Science and Engineering, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of SciencesShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityNational Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute, Chinese Academy of ScienceShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityKey Laboratory of RNA Innovation, Science and Engineering, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of SciencesShanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine, Rui-Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityAbstract Promyelocytic leukemia protein (PML) nuclear bodies (NBs) are essential in regulating tumor suppression, antiviral response, inflammation, metabolism, aging, and other important life processes. The re-assembly of PML NBs might lead to an ~100% cure of acute promyelocytic leukemia. However, until now, the molecular mechanism underpinning PML NB biogenesis remains elusive due to the lack of structural information. In this study, we present the cryo-electron microscopy (cryo-EM) structure of the PML dimer at an overall resolution of 5.3 Å, encompassing the RING, B-box1/2 and part of the coiled-coil (RBCC) domains. The integrated approach, combining crosslinking and mass spectrometry (XL-MS) and functional analyses, enabled us to observe a unique folding event within the RBCC domains. The RING and B-box1/2 domains fold around the α3 helix, and the α6 helix serves as a pivotal interface for PML dimerization. More importantly, further characterizations of the cryo-EM structure in conjugation with AlphaFold2 prediction, XL-MS, and NB formation assays, help unveil an unprecedented octopus-like mechanism in NB assembly, wherein each CC helix of a PML dimer (PML dimer A) interacts with a CC helix from a neighboring PML dimer (PML dimer B) in an anti-parallel configuration, ultimately leading to the formation of a 2 µm membrane-less subcellular organelle.https://doi.org/10.1038/s41421-024-00735-3
spellingShingle Yangxia Tan
Jiawei Li
Shiyan Zhang
Yonglei Zhang
Zhiyi Zhuo
Xiaodan Ma
Yue Yin
Yanling Jiang
Yao Cong
Guoyu Meng
Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
Cell Discovery
title Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
title_full Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
title_fullStr Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
title_full_unstemmed Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
title_short Cryo-EM structure of PML RBCC dimer reveals CC-mediated octopus-like nuclear body assembly mechanism
title_sort cryo em structure of pml rbcc dimer reveals cc mediated octopus like nuclear body assembly mechanism
url https://doi.org/10.1038/s41421-024-00735-3
work_keys_str_mv AT yangxiatan cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT jiaweili cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT shiyanzhang cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT yongleizhang cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT zhiyizhuo cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT xiaodanma cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT yueyin cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT yanlingjiang cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT yaocong cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism
AT guoyumeng cryoemstructureofpmlrbccdimerrevealsccmediatedoctopuslikenuclearbodyassemblymechanism